CN104083556A - Method for extracting konjak ceramide from konjak - Google Patents

Method for extracting konjak ceramide from konjak Download PDF

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Publication number
CN104083556A
CN104083556A CN201410336065.3A CN201410336065A CN104083556A CN 104083556 A CN104083556 A CN 104083556A CN 201410336065 A CN201410336065 A CN 201410336065A CN 104083556 A CN104083556 A CN 104083556A
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extraction
pressure
separating still
rhizoma amorphophalli
kettle
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CN104083556B (en
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葛发欢
李海池
史庆龙
马晋芳
张湘东
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Sun Yat Sen University
Guangzhou Zhongda Nansha Technology Innovation Industrial Park Co Ltd
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Sun Yat Sen University
Guangzhou Zhongda Nansha Technology Innovation Industrial Park Co Ltd
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    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P20/00—Technologies relating to chemical industry
    • Y02P20/50—Improvements relating to the production of bulk chemicals
    • Y02P20/54—Improvements relating to the production of bulk chemicals using solvents, e.g. supercritical solvents or ionic liquids

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Abstract

The invention discloses a method for extracting konjak ceramide from konjak. The method comprises the following steps: (1) crushing konjak into fine powder, adding ethanol, controlling the material-liquid ratio to be 1.5:(500-1200)Kg/ml, and granulating; (2) adding konjak grains prepared in the step (1) into an extraction kettle of supercritical CO2 extraction equipment, heating the extraction kettle, a separation kettle I and a separation kettle II, refrigerating a refrigerator and a storage tank, opening a carbon dioxide bottle when the temperature of the extraction kettle is 30-70 DEG C, the temperature of the separation kettle I is 40-65 DEG C and the temperature of the separation kettle II is 30-60 DEG C, pressurizing the extraction kettle and the separation kettle by a high-pressure pump, closing the carbon dioxide bottle when the pressure of the extraction kettle is 20-40MPa, the pressure of the separation kettle I is 6-18MPa and the pressure of the separation kettle II is 4-7MPa, opening a circulation valve for extracting circulation, discharging from the separation kettles I and II respectively every 30 minutes of circulation, and circularly extracting for 1-2 hours. The yield of C6-ceramide extracted by the method is high, and the extracted product has higher quality.

Description

A kind of method of extracting konjak ceramide from Rhizoma amorphophalli
Technical field
The present invention relates to extraction and separation technology field, relate in particular to a kind of method of extracting konjak ceramide from Rhizoma amorphophalli.
Background technology
Rhizoma amorphophalli (konjac) belongs to the herbal tuber of Araeceae (Araceae) Amorphophallus (Amorphophallus B1.ex Decne.), have another name called mill taro, terrible taro, ghost head, flower lotus bar, Rhizoma Amorphophalli etc., mainly originate in the torrid zone, the Eastern Hemisphere, subtropical zone, in China, have more widely and distribute.Rhizoma amorphophalli is cold in nature, pungent, poisonous; Can blood circulation promoting and blood stasis dispelling, removing toxic substances and promoting subsidence of swelling, wide intestinal relieving constipation, resolving phlegm and softening hard masses; Cure mainly the diseases such as blood pressure lowering, blood sugar lowering, scrofula sucutaneous nodule, damage congestive edema, constipation stomachache, laryngopharynx swelling and pain, gingival swelling and pain.
Ceramide type compound (Ceramide), N-fatty acyl-sphingosine, is the compounds being formed by sphingosine (sphingosine) and long-chain fatty acid condensation, is most important a kind of in sphingolipid compound.Ceramide is emerging a kind of active component recent years, contains than more rich ceramide in Rhizoma amorphophalli, and the content of its ceramide, up to 0.15～0.2%, is that currently used raw material is as 10 times of the raw materials such as Testa oryzae, Semen sojae atricolor, Semen Tritici aestivi.
What extraction ceramide was the most frequently used is organic solvent extraction, conventionally adopts isopropyl alcohol, ethyl acetate or chloroform extraction.Liu Renping etc., when exploring reflux extraction extraction konjak ceramide, find that the extraction effect of medium polar solvent ethyl acetate is the most desirable under the same conditions.But, under identical extraction conditions, with 95% ethanol extraction, by the effect of petroleum ether extraction and the extraction effect of ethyl acetate, be more or less the same, and ethyl acetate price is higher, in leaching process, volatilize larger, therefore, adopt 95% ethanol as extracting solvent.Author determines that by orthogonal test the optimised process extracting is 95% ethanol, extracts 80 ℃ of temperature, extraction time 8h, solid-liquid ratio 1:10, and the highest extracted amount reaches 4.34mg/g.
Urge splendid sunshine etc., adopt 95% ethanol, solid-liquid ratio 1:7.5,80 ℃～90 ℃ of bath temperatures, extraction time 8h～10h, adjusts extracting solution ethanol solubility to 70% with deionized water, petroleum ether extraction, Rotary Evaporators is concentrated.Concentrated solution is with petroleum ether: acetone (7:3) 200ml carries out 2 separation and purification of silica gel column chromatography, flow velocity is 1d/s, eluent is concentrated with Rotary Evaporators, concentrated solution carries out vacuum lyophilization and obtains white crude product, crude product carries out purification with acetone recrystallization, obtains the ceramide sample of white purification.
Wait quietly Wei Different Extraction Method to analyze mensuration, result shows that the content that solvent extraction method is measured under similar condition is 1.93%, and the content that supercritical ultrasonics technology records is 2.24%, and the latter's extraction effect is obviously better than the former.Author adopts experiments of single factor to analyze the principal element of the amide extraction ratio that affects the nerves, and utilizes orthogonal experiment to optimize the optimum process condition that extracts ceramide.Research shows, the optimum condition of supersound extraction konjak ceramide is: concentration of alcohol 95%, feed liquid mass ratio is 1:4, extracts temperature 60 C, extraction time 35min.Under this condition, the extraction ratio of ceramide crude extract is 2.89%.
Though the application that supercritical extraction technique extracts in ceramide at Rhizoma amorphophalli there is not yet report, Li Jia has applied this technology in microorganisms ferment rice bran is prepared ceramide, is indicating that supercritical extraction technique will extract ceramide at Rhizoma amorphophalli and be applied.Li Jia utilizes sphingomyelinase to make the content of ceramide free in Testa oryzae improve more than 3 times, and take extracting pressure as 30MPa, 32 ℃～35 ℃ of extraction temperature, separating still pressure is 7MPa～8MPa, the condition that temperature is 35 ℃～38 ℃ is carried out supercritical extraction and is obtained best effect of extracting.
Konjak ceramide is plant source ceramide, and safe in animal sources ceramide and synthetic ceramide, content is also originated higher than other plant.From existing document, the research of konjak ceramide is mainly concentrated on to its pharmacologically active aspect, and it extracts separation and quality analysis aspect is less, and extraction separation method tradition is loaded down with trivial details, the inadequate system of analytical method is comprehensive, also has no the supercritical CO of its ceramide 2extraction process research.
Summary of the invention
The invention provides a kind of method of extracting konjak ceramide from Rhizoma amorphophalli.
The present invention adopts following technical scheme:
The concrete steps of the method for extracting konjak ceramide from Rhizoma amorphophalli of the present invention are as follows:
(1) Rhizoma amorphophalli powder is broken into fine powder, then adds ethanol, solid-liquid ratio is 1.5:500-1200Kg/ml, then granulates;
(2) the Rhizoma amorphophalli granule of being prepared by step (1) adds supercritical CO 2in the extraction kettle of extraction equipment, to extraction kettle, separating still I, separating still II heats, and to cold machine and storage tank refrigeration, when extraction kettle temperature is 30-70 ℃, separating still I temperature is 40-65 ℃, when separating still II temperature is 30-60 ℃, open dioxide bottle, by high-pressure pump to extraction kettle, separating still pressurizes, when extraction kettle pressure is 20-40MPa, separating still I pressure is 6-18MPa, when separating still II pressure is 4-7MPa, closing carbon dioxide, pressure opening circulating valve, carry out extraction cycle, every circulation 30min, respectively from separating still I, II blowing, cycling extraction 1-2h.
In step (1), the concentration of ethanol is 95%, and solid-liquid ratio is preferably 1.5:1000Kg/ml.
In step (2), preferably extraction kettle temperature is 60 ℃, and pressure is 35MPa.
In step (2), preferable separate still I temperature is 50 ℃, and pressure is 15MPa.
In step (2), preferable separate still II temperature is 45 ℃, and pressure is 6MPa.
In step (2), preferably extraction time is 1.5h.
Technique effect of the present invention:
The method of extracting konjak ceramide from Rhizoma amorphophalli of the present invention is compared with solvent refluxing extraction method, supercritical CO 2extraction is compared with the C of reflux, extract, 6the yield of-ceramide is high by approximately 20%, and supercritical CO 2extraction product quality is better, at outward appearance, C 6the aspects such as-ceramide content are all better than solvent refluxing extraction method, and extraction process is simple simultaneously, efficiency is high, has solved the problems such as traditional reflux extraction solvent load is large, inflammable and explosive.
Accompanying drawing explanation
Fig. 1 is supercritical CO of the present invention 2extraction equipment schematic flow sheet.
In figure: 1-extraction kettle, 2-separating still I, 3-separating still II, 4-detached dowel, 5-heat exchanger, 6-high-pressure pump, 7-CO 2gas cylinder, the cold machine storage tank of 8-, 9-effusion meter.
The specific embodiment
The following examples are to describe in further detail of the present invention.
Embodiment 1
(1) Rhizoma amorphophalli powder is broken into fine powder, then adds ethanol, solid-liquid ratio is 1.5:500Kg/ml, then granulates;
(2) the Rhizoma amorphophalli granule of being prepared by step (1) adds supercritical CO 2in the extraction kettle of extraction equipment, extraction kettle, separating still I, separating still II are heated, and to cold machine and storage tank refrigeration, when extraction kettle temperature is 30 ℃, separating still I temperature is 40 ℃, when separating still II temperature is 30 ℃, opens dioxide bottle, by high-pressure pump, extraction kettle, separating still are pressurizeed, when extraction kettle pressure is 20MPa, separating still I pressure is 6MPa, when separating still II pressure is 4MPa, closing carbon dioxide, pressure opening circulating valve, carries out extraction cycle, every circulation 30min, respectively from separating still I, II blowing, cycling extraction 2h.
In step (1), the concentration of ethanol is 95%.
Embodiment 2
(1) Rhizoma amorphophalli powder is broken into fine powder, then adds ethanol, solid-liquid ratio is 1.5:1200Kg/ml, then granulates;
(2) the Rhizoma amorphophalli granule of being prepared by step (1) adds supercritical CO 2in the extraction kettle of extraction equipment, extraction kettle, separating still I, separating still II are heated, and to cold machine and storage tank refrigeration, when extraction kettle temperature is 70 ℃, separating still I temperature is 65 ℃, when separating still II temperature is 60 ℃, opens dioxide bottle, by high-pressure pump, extraction kettle, separating still are pressurizeed, when extraction kettle pressure is 40MPa, separating still I pressure is 18MPa, when separating still II pressure is 7MPa, closing carbon dioxide, pressure opening circulating valve, carries out extraction cycle, every circulation 30min, respectively from separating still I, II blowing, cycling extraction 1h.
In step (1), the concentration of ethanol is 95%.
Embodiment 3
(1) Rhizoma amorphophalli powder is broken into fine powder, then adds ethanol, solid-liquid ratio is 1.5:1000Kg/ml, then granulates;
(2) the Rhizoma amorphophalli granule of being prepared by step (1) adds supercritical CO 2in the extraction kettle of extraction equipment, extraction kettle, separating still I, separating still II are heated, and to cold machine and storage tank refrigeration, when extraction kettle temperature is 60 ℃, separating still I temperature is 50 ℃, when separating still II temperature is 45 ℃, opens dioxide bottle, by high-pressure pump, extraction kettle, separating still are pressurizeed, when extraction kettle pressure is 35MPa, separating still I pressure is 15MPa, when separating still II pressure is 6MPa, closing carbon dioxide, pressure opening circulating valve, carries out extraction cycle, every circulation 30min, respectively from separating still I, II blowing, cycling extraction 1.5h.
In step (1), the concentration of ethanol is 95%.
Comparative example (conventional solvent extraction)
Take Rhizoma amorphophalli powder, put into round-bottomed flask, add 95% ethanol, 80 ℃ of water-bath reflux, extract, 2 times, each 2h (adds 960ml95% ethanol for the first time, add for the second time 720ml95% ethanol), filter, merge extractive liquid,, 50 ℃ of concentrated by rotary evaporations are to anhydrous, obtain sepia oil-like extracts, weigh and calculated yield.
The calculating of yield
Yield is the important indicator of evaluating effect of extracting, and the computing formula of yield is as follows:
Crude extract yield=medical material crude extract quality ÷ medical material inventory * 100%
C6-Cer percentage composition % ÷ medical material inventory * 100% in C6-Cer yield=medical material crude extract quality * crude extract.
The yield of embodiment 1-3 is as shown in table 1:
Table 1
Take embodiment 3 as example contrast supercritical CO 2the quality that extraction and conventional solvent are extracted, result is as shown in table 2:
Table 2 supercritical CO 2the comparison of extraction and traditional reflux extraction
As can be seen from Table 2: supercritical CO 2extraction konjak ceramide crude extract yield is lower than reflux extraction, and C 6-Cer yield is higher, may contain the impurity large compared with pluripolarity due to the product of solvent extraction.Compare with traditional ethanol reflux extraction, supercritical extract is all better than traditional reflux extraction at aspects such as color and luster, ceramide content and yields.Therefore, adopt supercritical CO 2extraction konjak ceramide, is better than conventional extracting method.
Compare with conventional solvent reflux extraction, the supercritical CO of konjak ceramide of the present invention 2extraction process has following characteristics:
(1) extracting power is strong, and extraction efficiency is high, and yield is higher by approximately 20% than reflux extraction;
(2) extraction temperature is low, effectively protects temperature-sensitive effective ingredient;
(3) energy selective extraction and separated component, separable impurity in the time of extraction process;
(4) production technology is simple, does not need the techniques such as filtration;
(5) extraction is carried out under high pressure inert conditions, almost can kill aerobic microbiological completely, effectively guarantees and improve the quality of product
(6) avoid the use of organic solvent, reduce the consumption of ethanol.
Rhizoma amorphophalli supercritical CO 2extract analysis of volatile components
Embodiment 3 volatile ingredient preparation and konjak ceramide extract prepared by solvent method is carried out to makings chromatographic comparison:
GC conditions: chromatographic column is TR-1MS type 100% dimethyl polysiroxan fused-silica capillary column (30m * 0.25mm * 0.25 μ m); 250 ℃ of injector temperatures, do not shunt, and carrier gas is helium, flow velocity 1ml/min, sample size: 1 μ L.Mass spectrum condition: 280 ℃ of transfer tube temperature; 230 ℃ of ion source temperatures; EI ion source, electron energy 70ev; Full scan, sweep limits: 40-450.
Respectively the product of embodiment 3, commercially available prod and comparative example is carried out to component analysis, result is as shown in table 3:
3 kinds of different product volatile ingredient contrast tables of table 3
Known according to table 3 analysis result, there is larger difference in composition kind and number in three kinds of samples, Rhizoma amorphophalli supercritical CO 2extract main component is the materials such as Ethyl linoleate, ethyl palmitate, bean Gona-4-en-3-one, linolenic acid, elaidic acid ethyl ester; Rhizoma amorphophalli commercially available prod main component is the materials such as methyl linoleate, chionasterol, stigmasterol, vitamin E; Rhizoma amorphophalli reflux, extract, thing main component is diisobutyl phthalate, chionasterol, stigmasterol, 2,6-dimethyl-1,6-glycol-2, the materials such as 7-octadiene.
Rhizoma amorphophalli supercritical CO 2fatty acid in extract (ester) class material is mainly the compositions such as linolenic acid, Ethyl linoleate, ethyl palmitate, elaidic acid ethyl ester, and total relative amount reaches 71.68%; Rhizoma amorphophalli reflux, extract, material does not almost have this type of material; Commercially available Rhizoma amorphophalli extract mainly contains the compositions such as methyl linoleate, ethyl palmitate, elaidic acid ethyl ester, and relative amount is 31.61%.
Rhizoma amorphophalli supercritical CO 2in extract, main sterol is the materials such as bean Gona-4-en-3-one, bean sterone, 4-cholestene-3-ketone, and total relative amount reaches 12.93%; It is chionasterol, stigmasterol and glycerol deoxycholic acid that Rhizoma amorphophalli reflux, extract, owner wants sterol, and relative amount is 18.21%; Commercially available Rhizoma amorphophalli extract is mainly containing chionasterol and stigmasterol, and relative amount is 14.31%.
Squalene is a kind of lipid non-saponifiable matter.In three kinds of Rhizoma amorphophalli extracts, all contain Squalene, at supercritical CO 2relative amount in extract is 1.71%, is 1.84% in reflux, extract, thing, is 3.39% in commercially available extract.Squalene can promote hepatocellular regeneration and protect hepatocyte, thereby improve the function of liver; Squalene can impel the interior Superoxide dismutase of body and lactic acid dehydrogenase significantly to raise, and promotes lactic acid to decompose, and makes energy i (in vivo) metabolism vigorous, the fast quick-recovery of muscle power, timely allaying tiredness; Squalene energy blood circulation promoting, the content of reduction cholesterol and triglyceride, suppresses cholesterol in serum concentration, reduces lipoprotein concentration, and accelerates the discharge of cholesterol, can delay atherosclerotic formation, the generation of prevention cardiovascular and cerebrovascular disease.In addition, Squalene also has the effects such as defying age, antitumor.
Vitamin E is a kind of fatsoluble vitamin, claims again tocopherol, is one of topmost antioxidant.Relative amount in Rhizoma amorphophalli supercritical extract is 0.63%, is 6.00% in commercially available prod, and in reflux, extract, thing, quality testing is measured.Vitamin E has physiological function widely, and it can be by self being oxidized to fertility quinone, by active ROO ﹣be transformed into inactive ROOH, thereby interrupt the chain reaction of lipid peroxidation, suppress the peroxidation of lipid, Cell protection escapes injury; Promote sex hormones secretion, man's motility of sperm and quantity are increased, improve fertility; Maintain the permeability of blood capillary, increase blood flow, strengthen the ability of the cold resisting, and repair the cicatrix after vascular damaged, suppress hematoblastic gathering, the formation of anti-tampon.The Alcohol soluble composition major part of Rhizoma amorphophalli has physiological activity, identical or close with the part effect of Rhizoma amorphophalli, can infer that the part effect of Rhizoma amorphophalli may be relevant with its Alcohol soluble composition.For the further development and utilization of Rhizoma amorphophalli provides the foundation of science.
Although illustrated and described embodiments of the invention, for the ordinary skill in the art, be appreciated that without departing from the principles and spirit of the present invention and can carry out multiple variation, modification, replacement and modification to these embodiment, scope of the present invention is limited by claims and equivalent thereof.

Claims (6)

1. from Rhizoma amorphophalli, extract a method for konjak ceramide, it is characterized in that: the concrete steps of described method are as follows:
(1) Rhizoma amorphophalli powder is broken into fine powder, then adds ethanol, solid-liquid ratio is 1.5:500-1200Kg/ml, then granulates;
(2) the Rhizoma amorphophalli granule of being prepared by step (1) adds supercritical CO 2in the extraction kettle of extraction equipment, to extraction kettle, separating still I, separating still II heats, and to cold machine and storage tank refrigeration, when extraction kettle temperature is 30-70 ℃, separating still I temperature is 40-65 ℃, when separating still II temperature is 30-60 ℃, open dioxide bottle, by high-pressure pump to extraction kettle, separating still pressurizes, when extraction kettle pressure is 20-40MPa, separating still I pressure is 6-18MPa, when separating still II pressure is 4-7MPa, closing carbon dioxide, pressure opening circulating valve, carry out extraction cycle, every circulation 30min, respectively from separating still I, II blowing, cycling extraction 1-2h.
2. the method for extracting konjak ceramide from Rhizoma amorphophalli as claimed in claim 1, is characterized in that: in step (1), the concentration of ethanol is 95%, and solid-liquid ratio is 1.5:1000Kg/ml.
3. the method for extracting konjak ceramide from Rhizoma amorphophalli as claimed in claim 1, is characterized in that: in step (2), extraction kettle temperature is 60 ℃, and pressure is 35MPa.
4. the method for extracting konjak ceramide from Rhizoma amorphophalli as claimed in claim 1, is characterized in that: in step (2), separating still I temperature is 50 ℃, and pressure is 15MPa.
5. the method for extracting konjak ceramide from Rhizoma amorphophalli as claimed in claim 1, is characterized in that: in step (2), separating still II temperature is 45 ℃, and pressure is 6MPa.
6. the method for extracting konjak ceramide from Rhizoma amorphophalli as claimed in claim 1, is characterized in that: in step (2), extraction time is 1.5h.
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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105403633A (en) * 2015-10-28 2016-03-16 广州中大南沙科技创新产业园有限公司 Method for determining content of ceramide in konjak
CN107721871A (en) * 2017-10-20 2018-02-23 中山大学 A kind of method that molecular distillation isolates and purifies konjak ceramide
CN113413632A (en) * 2021-07-08 2021-09-21 宁夏大学 A lipid extract of Stellaria dichotoma and its preparation method

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CN103755583A (en) * 2014-01-27 2014-04-30 广州市娇兰化妆品有限公司 Method for extracting and separating ceramide in konjac flying powder

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CN103755583A (en) * 2014-01-27 2014-04-30 广州市娇兰化妆品有限公司 Method for extracting and separating ceramide in konjac flying powder

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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105403633A (en) * 2015-10-28 2016-03-16 广州中大南沙科技创新产业园有限公司 Method for determining content of ceramide in konjak
CN105403633B (en) * 2015-10-28 2018-05-11 广州中大南沙科技创新产业园有限公司 Ceramide content assay method in konjaku
CN107721871A (en) * 2017-10-20 2018-02-23 中山大学 A kind of method that molecular distillation isolates and purifies konjak ceramide
CN113413632A (en) * 2021-07-08 2021-09-21 宁夏大学 A lipid extract of Stellaria dichotoma and its preparation method
CN113413632B (en) * 2021-07-08 2022-06-03 宁夏大学 A lipid extract of Stellaria dichotoma and its preparation method

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